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. 2005 Oct;17(10):2693–2704. doi: 10.1105/tpc.105.034959

Figure 1.

Figure 1.

Characterization of the IAA Carboxyl Methyltransferase Gene IAMT1.

(A) to (C) Top, 26-d-old wild-type, heterozygous, and homozygous IAA carboxyl methyltransferase1-dominant (iamt1-D) mutant Arabidopsis plants. Bottom, leaf transverse sections from the corresponding plants.

(D) Scheme of the genomic region flanking the T-DNA insertion site in iamt1-D. Genes are represented by different colored arrows, intergenic regions by lines, and the T-DNA insertion site by a black X. The arrow direction represents the transcriptional orientation of the genes. The four red arrowheads represent the four 35S enhancers from pSKI015. The thin black arrows represent the primers used in cosegregation analysis. LB, T-DNA left border; bar, Basta resistance gene; 4Enhancer, CaMV 35S enhancer tetrad; RB, T-DNA right border.

(E) Linkage analysis of the T-DNA insertion and hyponastic phenotypes. P1 and P2 will amplify a 357-bp fragment from the wild type, and P1 and LS4 will amplify a 136-bp fragment from the homozygous iamt1-D mutant. Marker, 100-bp ladder.

(F) Expression of IAMT1 in the wild type and the homozygous iamt1-D mutant by RT-PCR with the primer pair IAMT-1 and IAMT-2. TUB2, β-tubulin gene as an internal control.

(G) Recapitulation of iamt1-D phenotypes by overexpression of IAMT1 cDNA. The IAMT1 cDNA (coding region) was overexpressed under the control of four copies of the 35S enhancers and the IAMT1 promoter. The transgenic plants displayed hyponastic leaf phenotypes. LB, T-DNA left border; polyA, CaMV 35S poly(A); KanR, kanamycin resistance gene NPTII; 35S-P, CaMV 35S promoter; 4Enhancer, CaMV 35S enhancer tetrad; IAMT1-P, promoter of IAMT1; IAMT1, open reading frame of IAMT1; Ter, nopaline synthase terminator; RB, T-DNA right border.

(H) Construct of the IAMT1-RNAi plasmid. LB, polyA, KanR, 35S-P, IAMT1, Ter, and RB are as in (G). GUSF, 1-kb fragment of GUS.

(I) Suppression of iamt1-D phenotypes by the IAMT1-RNAi construct. A 26-d-old iamt1-D homozygous mutant plant transformed with the IAMT1-RNAi construct is shown. Bar = 1 mm.